Skip to main content
. 2013 Jun;87(11):6482–6491. doi: 10.1128/JVI.03428-12

Fig 5.

Fig 5

Interaction of Hsp90 with the BALF5 DNA polymerase catalytic subunit. (A) Coimmunoprecipitation of Hsp90 with BALF5. HEK293T cells were transfected with pcDNA-FLAG-BALF5 alone, pcDNA-HA-Hsp90 alone, or both pcDNA-FLAG-BALF5 and pcDNA-HA-Hsp90. Cells were harvested at 24 h posttransfection, and immunoprecipitation (IP) analysis was carried out as described in Materials and Methods. The antibodies used for IP and subsequent immunoblot (IB) analysis are indicated. (B) Disruption of the interaction between BALF5 and Hsp90 by radicicol. HEK293T cells were transfected with pcDNA-FLAG-BALF5 and pcDNA-HA-Hsp90 in the absence or the presence of 0.5 μM radicicol. Cells were harvested at 24 h posttransfection, and IP was carried out, as described in Materials and Methods. Antibodies used for IP and subsequent IB are indicated. (C) The indicated amounts of input samples of panel B were electrophoresed and analyzed by immunoblotting with anti-FLAG, anti-cdc2, and antitubulin antibodies. (D) Stabilization of the interaction between BALF5 and Hsp90 by molybdate. HEK293T cells transfected with pcDNA-FLAG-BALF5 and pcDNA-HA-Hsp90 were lysed in 0.5% mCSK buffer in the presence or absence of 20 mM molybdate. Lysates were subjected to IP and subsequent IB analyses. The antibodies used for IP and IB are indicated. (E) Hsp90 does not associate with BMRF1. HEK293T cells were transfected with pcDNA-BMRF1 and pcDNA-HA-Hsp90 in the absence or presence of 20 mM molybdate. Cells were harvested at 24 h posttransfection, and IP was carried out as described in Materials and Methods. The antibodies used for IP and subsequent IB analyses are indicated. (F) Hsp90 contributes to BALF5-BMRF1 complex formation. HEK293T cells were transfected with pcDNA-FLAG-BALF5, pcDNA-HA-Hsp90, and pcDNA-BMRF1. Cells were harvested at 24 h posttransfection, and IP was carried out as described in Materials and Methods. The antibodies used for IP and subsequent IB analyses are indicated. On each panel, the input lane contains 10% of the amount of the soluble fractions that were subjected to immunoprecipitation.